Characterization of two thioredoxins h with predominant localization in the nucleus of aleurone and scutellum cells of germinating wheat seeds

Characterization of two thioredoxins h with predominant localization in the nucleus of aleurone... Two full-length cDNA clones, designated TrxhA and TrxhB, encoding different but very similar thioredoxin h polypeptides were isolated from wheat (Triticum aestivum cv. Chinese Spring) aleurone cells. The deduced proteins show a high similarity to each other and to thioredoxin h from other sources, in particular from T. aestivum and T. durum. One of them, TRXhA, was expressed in E. coli as a His-tagged polypeptide and used to raise polyclonal antibodies by immunization of rabbits. These antibodies identified a single band (ca. 13.5 kDa) in western blot analysis of protein extracts from all wheat organs analyzed. TRXhA and TRXhB when expressed in E. coli as intact polypetides showed indistinguishable electrophoretic mobility, which corresponded to the 13.5 kDa polypeptide detected in wheat protein extracts. The amount of thioredoxin h transcripts increased in scutellum and aleurone cells during germination but GA3 did not exert any stimulatory effect on thioredoxin h expression. Although northern blot analysis detected a single band, competitive RT-PCR showed that this band is due to the accumulation of both TrxhA and TrxhB mRNAs. These results suggest that the single band detected in western blots is due to the presence of at least two thioredoxin h polypeptides. Immunolocalization experiments confirmed the high content of thioredoxins h in scutellum and aleurone cells, and showed a low content in the starchy endosperm of germinating grains. Interestingly, though these proteins are evenly distributed in the cytosol, the highest levels of thioredoxins h were detected in the nucleus, both in aleurone and scutellum cells. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Molecular Biology Springer Journals

Characterization of two thioredoxins h with predominant localization in the nucleus of aleurone and scutellum cells of germinating wheat seeds

Loading next page...
 
/lp/springer_journal/characterization-of-two-thioredoxins-h-with-predominant-localization-Veug0ffKHB
Publisher
Kluwer Academic Publishers
Copyright
Copyright © 2001 by Kluwer Academic Publishers
Subject
Life Sciences; Biochemistry, general; Plant Sciences; Plant Pathology
ISSN
0167-4412
eISSN
1573-5028
D.O.I.
10.1023/A:1010697331184
Publisher site
See Article on Publisher Site

Abstract

Two full-length cDNA clones, designated TrxhA and TrxhB, encoding different but very similar thioredoxin h polypeptides were isolated from wheat (Triticum aestivum cv. Chinese Spring) aleurone cells. The deduced proteins show a high similarity to each other and to thioredoxin h from other sources, in particular from T. aestivum and T. durum. One of them, TRXhA, was expressed in E. coli as a His-tagged polypeptide and used to raise polyclonal antibodies by immunization of rabbits. These antibodies identified a single band (ca. 13.5 kDa) in western blot analysis of protein extracts from all wheat organs analyzed. TRXhA and TRXhB when expressed in E. coli as intact polypetides showed indistinguishable electrophoretic mobility, which corresponded to the 13.5 kDa polypeptide detected in wheat protein extracts. The amount of thioredoxin h transcripts increased in scutellum and aleurone cells during germination but GA3 did not exert any stimulatory effect on thioredoxin h expression. Although northern blot analysis detected a single band, competitive RT-PCR showed that this band is due to the accumulation of both TrxhA and TrxhB mRNAs. These results suggest that the single band detected in western blots is due to the presence of at least two thioredoxin h polypeptides. Immunolocalization experiments confirmed the high content of thioredoxins h in scutellum and aleurone cells, and showed a low content in the starchy endosperm of germinating grains. Interestingly, though these proteins are evenly distributed in the cytosol, the highest levels of thioredoxins h were detected in the nucleus, both in aleurone and scutellum cells.

Journal

Plant Molecular BiologySpringer Journals

Published: Oct 3, 2004

References

You’re reading a free preview. Subscribe to read the entire article.


DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 18 million articles from more than
15,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

Search

Query the DeepDyve database, plus search all of PubMed and Google Scholar seamlessly

Organize

Save any article or search result from DeepDyve, PubMed, and Google Scholar... all in one place.

Access

Get unlimited, online access to over 18 million full-text articles from more than 15,000 scientific journals.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

DeepDyve

Freelancer

DeepDyve

Pro

Price

FREE

$49/month
$360/year

Save searches from
Google Scholar,
PubMed

Create lists to
organize your research

Export lists, citations

Read DeepDyve articles

Abstract access only

Unlimited access to over
18 million full-text articles

Print

20 pages / month

PDF Discount

20% off